CN112891363A

CN112891363A - New use of New Zealand Vitexin 2 and New Zealand Vitexin 3

Info

Publication number: CN112891363A
Application number: CN202110316466.2A
Authority: CN
Inventors: 孙铭飞; 廖申权; 戚南山; 林栩慧; 吕敏娜; 吴彩艳; 李娟�; 蔡海明; 胡俊菁; 肖文婉; 于林增; 张小慧; 张健騑; 谢明权
Original assignee: Institute of Animal Health of Guangdong Academy of Agricultural Sciences
Current assignee: Institute of Animal Health of Guangdong Academy of Agricultural Sciences
Priority date: 2021-03-23
Filing date: 2021-03-23
Publication date: 2021-06-04
Anticipated expiration: 2041-03-23
Also published as: CN112891363B

Abstract

The invention relates to the technical field of animal disease prevention and control, in particular to a new application of New Zealand vitexin 2 and New Zealand vitexin 3, and specifically relates to an application of New Zealand vitexin 2 and New Zealand vitexin 3 in preparation of a coccidiosis-resistant medicament or feed. Compared with the prior art, the invention has the following beneficial effects: according to the invention, the new zealand vitexin 2 and new zealand vitexin 3 have obvious inhibition effects on coccidia, the inhibition effects are embodied in that the new zealand vitexin 2 and new zealand vitexin 3 can effectively inhibit the propagation of eimeria coccidia on a cell culture model of the eimeria coccidia, and the inhibition effects are verified through animal body tests. Aiming at the discovery, the application fields of the New Zealand vitexin 2 and the New Zealand vitexin 3 are widened, and a new scheme is provided for the development of anti-coccidial drugs.

Description

New use of New Zealand Vitexin 2 and New Zealand Vitexin 3

Technical Field

The invention relates to the technical field of animal disease prevention and control, in particular to new application of New Zealand vitexin 2 and New Zealand vitexin 3.

Background

New Zealand Vitexin 2, also known as Wei-tannin-2, has molecular weight of 594.5, CAS number of 23666-13-9, and molecular formula of C₂₇H₃₀O₁₅The chemical structural formula is shown as follows:

new Zealand vitexin 3, also known as visanin-3, has molecular weight of 564.49,CAS number 59914-91-9, molecular formula C₂₆H₂₈O₁₄The chemical structural formula is shown as follows:

new Zealand vitexin 2 and new Zealand vitexin 3 are flavonoid glycoside compounds, and are distributed in leaves, stems, roots, barks, fruits and seeds of plants. The existing research shows that the New Zealand vitexin 2 and the New Zealand vitexin 3 have the effects of oxidation resistance, virus resistance and inflammation resistance. According to the report, the functions of the New Zealand vitexin 2 and the New Zealand vitexin 3 comprise an anti-tumor effect and a myocardial protection effect.

Coccidiosis in chickens is a disease caused by infection with one or more species of eimeria, which are protozoa of the phylum apicomplexa, class sporozoea, order coccidioideae, family eimeria, genus eimeria. There are 7 species of Eimeria tenella (Eimeria tenella), Eimeria necatrix (e.necatrix), Eimeria brunetti (e.brunetti), Eimeria acervulina (e.acervulina), Eimeria maxima (e.maxima), Eimeria mitis (e.mitis) and Eimeria praecox (e.praecox), respectively.

Eimeria causing coccidiosis in chickens mainly includes eimeria tenella (e.tenella), eimeria necatrix (e.necatrix). After the chicken only infected with the Eimeria, the production performance is seriously reduced, even death is caused, and serious economic loss is brought to the chicken industry. Eimeria tenella (e.tenella) is one of the most harmful species, and after infection of a chicken, the chicken parasitizes in the caecum, resulting in clinical symptoms of bloody stool, anorexia, weight gain reduction and the like.

At present, the prevention and control of chicken coccidiosis at home and abroad mainly depends on medicines, the used medicines comprise two types of chemically synthesized medicines and antibiotics, and since the first appearance of special anticoccidial medicines in 1936, more than 40 anticoccidial medicines are reported, and 20 anticoccidial medicines are widely used at present. The traditional preventive drugs include proguanil, amproline, dinitolmide, monensin, salinomycin, diclazuril, maduramycin, clopyralid, nicarbazin, and the like. The traditional therapeutic drugs include toltrazuril solution, sulfonamides (such as sulfaquinoxaline and sulfaclozine sodium) and the like.

The prevention and treatment of the chicken coccidiosis depend on the use of the anticoccidial drugs, and the effect of the original anticoccidial drugs is obviously reduced along with the generation of the drug resistance of the clinical wild toxid strains to the traditional anticoccidial drugs, so that the anticoccidial drugs with definite treatment effects are lacking clinically. Therefore, a new anti-coccidiosis drug is urgently needed, and with the development needs of animal husbandry aquaculture and public health safety, the market demand for the new anti-coccidiosis drug will increase.

Disclosure of Invention

Based on the above, the main purpose of the invention is to provide a new application of the New Zealand vitexin 2 and the New Zealand vitexin 3, in particular to an application of the New Zealand vitexin 2 and the New Zealand vitexin 3 in preparing a medicine or feed for resisting coccidiosis, wherein the New Zealand vitexin 2 and the New Zealand vitexin 3 can effectively inhibit the breeding of coccidiosis and have obvious effect of resisting coccidiosis.

The purpose of the invention is realized by the following technical scheme:

use of at least one of New Zealand vitexin 2 and New Zealand vitexin 3 in preparing medicine or feed for resisting coccidiosis.

In one embodiment, the coccidiosis is chicken coccidiosis.

In one embodiment, the infectious protozoan of chicken coccidiosis is eimeria tenella.

In one embodiment, the chicken breed infected with the chicken coccidiosis is green south yellow broiler.

In one embodiment, the medicament comprises at least one of New Zealand vitexin 2 and New Zealand vitexin 3 and pharmaceutically acceptable auxiliary materials.

In one embodiment, the mode of administration of the drug is by mass administration.

In one embodiment, the medicament is in the form of a powder.

In one embodiment, the medicament is in the form of a water-soluble powder.

In one embodiment, the medicament is in the form of a solution.

In one embodiment, the drug is in the form of microcapsules.

In one embodiment, the dosage form of the drug is a tablet.

In one embodiment, the feed is a complete formula feed, a concentrated feed, a premix/premix feed, a concentrate mix, or a mixed feed.

Compared with the prior art, the application of at least one of the New Zealand vitexin 2 and the New Zealand vitexin 3 in preparing the anti-coccidiosis drug or feed has the following beneficial effects:

according to the invention, through research, the new zealand vitexin 2 and new zealand vitexin 3 have obvious inhibition effect on coccidia (especially on Eimeria tenella), the inhibition effect is embodied in that the new zealand vitexin 2 and the new zealand vitexin 3 can effectively inhibit the propagation of Eimeria on a culture model of Eimeria cells (such as MDBK cells), and the inhibition effect is verified through animal body tests. Aiming at the discovery, the application fields of the New Zealand vitexin 2 and the New Zealand vitexin 3 are developed, and a solution is provided for the development of novel anticoccidial drugs.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a graph showing the effect of Vitexin 2 in New Zealand on the reproduction of Eimeria tenella according to one embodiment of the invention; in the figure, the abscissa is the concentration of compound vitexin 2, and the ordinate is the inhibition rate of compound vitexin 2 against the propagation of eimeria tenella;

FIG. 2 is a graph showing the effect of Vitexin 3 in New Zealand on the reproduction of Eimeria tenella according to an embodiment of the invention; in the figure, the abscissa is the concentration of compound vitexin 3, and the ordinate is the inhibition rate of compound vitexin 3 against the propagation of eimeria tenella;

FIG. 3 is a graph showing the effect of sulfachloropyrazine sodium on the propagation of Eimeria tenella; in the figure, the abscissa is the concentration of the compound sulfachloropyrazine sodium, and the ordinate is the inhibition rate of the compound sulfachloropyrazine sodium on the propagation of Eimeria tenella.

Detailed Description

In order that the invention may be more fully understood, reference will now be made to the following description. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The embodiment of the invention provides application of at least one of New Zealand vitexin 2 and New Zealand vitexin 3 in preparation of a coccidiosis-resistant medicament or feed.

The embodiment of the invention discovers that the New Zealand vitexin 2 and the New Zealand vitexin 3 have obvious inhibition effect on coccidia (especially on Eimeria tenella), the inhibition effect is embodied in that the New Zealand vitexin 2 and the New Zealand vitexin 3 can effectively inhibit the propagation of Eimeria on a culture model of Eimeria cells (such as MDBK cells), and the inhibition effect is verified by animal body tests. Aiming at the discovery, the application fields of the New Zealand vitexin 2 and the New Zealand vitexin 3 are widened, and a new scheme is provided for the development of anti-coccidial drugs.

It will be appreciated that the "chicken coccidiosis" described in the examples of the present invention is a parasitic disease which can result in significant economic losses in poultry production. Eimeria tenella parasitizes in chicken cecal epithelial cells, causing intestinal tract tissue lesion, reducing feed intake and absorption rate of nutrients in feed, dehydration, bloody stool and the like. Coccidiosis in chickens is caused by infection with one or more species of Eimeria (Eimeria) coccidia.

It is to be understood that the "Drugs" described in the examples of the present invention are Veterinary Drugs, and the "Veterinary Drugs (Veterinary Drugs)" described in the examples of the present invention: means a substance (drug-containing feed additive) for preventing, treating, diagnosing animal diseases or purposefully regulating animal physiological functions.

The anti-coccidiosis in the embodiment of the invention can have an obvious treatment effect on the coccidiosis and also can have an obvious prevention effect on the coccidiosis. The therapeutic, prophylactic or preventative action includes, but is not limited to, inhibition of Eimeria's reproduction.

In a specific example, the coccidiosis is chicken coccidiosis.

In a specific example, the infectious protozoan of chicken coccidiosis is eimeria tenella.

In a specific example, the chicken breed infected with coccidiosis is green south yellow broiler.

In a specific example, the medicament comprises at least one of New Zealand vitexin 2 and New Zealand vitexin 3 and pharmaceutically acceptable auxiliary materials. It is understood that the type of the selected adjuvant, the amount of the selected adjuvant, etc. are different according to the dosage form of the drug.

It will be appreciated that the mode of administration of the medicament described in the examples of the invention is by mass administration. Of course, administration other than mass administration may be used, or a combination of mass administration with other pharmaceutically acceptable administration may be used.

It is understood that the "population administration" described in the examples of the present invention includes, but is not limited to:

mixing water for administration: first, the solubility of the drug in water is known; secondly, the dosage of the medicine is calculated according to the drinking amount, and if the stability of the medicine in water is poor, a thirst medicine taking method can be considered;

a material mixing and administration method: the drug is mixed into the feed for free ingestion, but the drug and the feed must be mixed evenly, and a progressive dilution method can be adopted.

In addition to the two ways of "group administration" of "mixed administration" and "mixed administration" as exemplified in the embodiments of the present invention, other suitable and veterinary acceptable ways of "group administration" may be adopted, and a plurality of ways of "group administration" may be used in combination.

It is understood that the New Zealand Vitex glycosides 2 and New Zealand Vitex glycosides 3 can be combined with suitable types of excipients to prepare different forms of drugs (e.g. liquid, semi-solid, solid) to form different dosage forms, including but not limited to: powder (including premix), water-soluble powder (including water-diffusible powder and water-soluble granule), solution (mainly aqueous solution, also suspension, emulsion, etc.), microcapsule, and tablet.

The water soluble powder (including water diffusible powder and water soluble granule) is made up by using medicine and auxiliary material of cosolvent and suspension adjuvant. Adding into drinking water to dissolve the medicine, and uniformly dispersing for animals to drink. The advantages of the dosage form include: the mixture is relatively uniform. ② the medicine has good effect for preventing and treating group diseases, especially sick livestock and poultry often eat less (even no) and drink more water, which is very good for guaranteeing that the sick livestock and poultry can eat enough medicine.

Solution (including oral liquid, suspension, and emulsion) is prepared by dissolving medicine in water, and can be used for oral administration or topical application. The advantages of the dosage form include: adding into drinking water to dissolve the medicine, and uniformly dispersing for animals to drink. The advantages of the dosage form include: the mixture is relatively uniform. ② the medicine has good effect for preventing and treating group diseases, especially sick livestock and poultry often eat less (even no) and drink more water, which is very good for guaranteeing that the sick livestock and poultry can eat enough medicine.

Powder, a widely used pharmaceutical dosage form, is ground into a powder form that can be incorporated into feed or dissolved in water for feeding to animals. The advantages of the dosage form include: the compound is added into feed as additive for preventing and treating diseases. Saves labor and time, can be used as clinical medicine to be mixed in the feed, and can also be directly used in the feed production process. ② convenient to store. And the quality is relatively stable.

Microcapsules are microcapsules which are prepared by wrapping a drug (core material) with a natural or synthetic polymer material (capsule material) and have a diameter of 1-500 microns. The medicinal microcapsule can be made into powder, tablet, injection, ointment, etc. according to requirement. The advantages of the dosage form include: the microcapsule can prolong drug effect, improve drug stability or cover unpleasant odor of the drug.

In addition to the forms of "powder", "water-soluble powder", "solution", "microcapsule" and "tablet", which are exemplified in the present embodiment, the new zealand vitexin 2 and the new zealand vitexin 3 can be prepared into other suitable and pharmaceutically acceptable veterinary drug forms. And it is understood that one dosage form or a combination of dosage forms of the New Zealand Vitexin 2 and New Zealand Vitexin 3 can be selected for feeding during the actual cultivation process.

The embodiment of the invention does not specially limit the specific types of the feed, and can be complete compound feed, concentrated feed, premix/premix feed, concentrated feed and mixed feed. The complete mixed feed, concentrated feed, premixed feed and concentrated feed mixture are mixed based on the growth stage, physiological requirement, nutritive requirement and nutritive value of animal, and are produced in certain proportion.

The complete compound feed is prepared from energy feed, protein feed, mineral feed, vitamins, amino acids and trace element additive according to a specified feeding standard, and has comprehensive and balanced nutrition.

A concentrated feed is prepared from protein feed, mineral feed (calcium, phosphorus and edible salt), and additive premix by mixing at a certain ratio. Can not be directly fed, and can be fed after corn or other energy feed is added.

The premixed feed is a mixture of one or more kinds of trace additive material, carrier and diluent, and its trace components are premixed to distribute homogeneously in great amount of feed. The feed is a semi-finished product, and can not be directly fed.

The concentrated feed mixture is a supplement concentrated feed for nutrient deficiency, mainly comprises energy feed, protein feed and mineral feed, and can be directly fed.

The mixed feed is prepared by simply processing and mixing certain feed raw materials, is a primary mixed feed, and mainly considers nutritional indexes such as energy, protein, calcium, phosphorus and the like.

The following will explain in detail the new uses of New Zealand Vitexin 2 and New Zealand Vitexin 3 according to the present invention with reference to specific examples. The starting materials used in the following examples are all commercially available products unless otherwise specified.

Example 1

This example provides a cellular level assay for New Zealand Vitexin 2 and New Zealand Vitexin 3, as follows:

firstly, experimental materials:

(1) medicine preparation: new Zealand Vitexin 2, New Zealand Vitexin 3 and sulfachloropyrazine sodium were purchased from Merck.

(2) Coccidian oocysts: eimeria tenella (e.tenella) eurycosporulated oocysts were stored by the institute of animal health, academy of agricultural sciences, guangdong, and rejuvenated in coccidiless chicks before use.

(3) Cell: MDBK cells (i.e. bovine kidney cells), were maintained by the institute of animal health, academy of agricultural sciences, guangdong.

II, an experimental method:

in this example, the e.tenella (e.tenella) resistance effect was evaluated by using an e.tenella MDBK cell culture model, and the specific evaluation steps included:

(1) MDBK cells grown to a confluency of 80% or more were digested with 0.25% trypsin digestion solution at 4X 10⁵Cell concentration per well plated 12 well cell culture plates at 37 ℃ with 5% CO₂The culture box is used for culturing for 24 hours.

(2) When the cells in the 12-well culture plate are grown to a confluent state of 80% or more, 10X 10 cells are inoculated per well⁴Sporozoites/ml, medium was discarded after 4h, washed 3 times with PBS, suspended sporozoites washed off and 5% fetal bovine serum in MEM medium was added to each well.

(3) Grouping tests:

setting 5 drug concentration gradients of a New Zealand vitexin 2 group, specifically 0.01 mu M, 0.1 mu M, 1 mu M, 10 mu M and 100 mu M, and setting 3 repetitions of each concentration;

setting 5 drug concentration gradients of a 3 group of New Zealand vitexin, specifically 0.01 mu M, 0.1 mu M, 1 mu M, 10 mu M and 100 mu M, and setting 3 repetitions of each concentration;

a positive control group (added with sulfachloropyrazine sodium) is provided with 5 drug concentration gradients, specifically 0.01 μ M, 0.1 μ M, 1 μ M, 10 μ M and 100 μ M, and each concentration is provided with 3 repetitions;

blank control group: adding volumes of vitexin 2, vitexin 3 and positive control drug into the new zealand vitexin 2 group, new zealand vitexin 3 group and positive control group, adding equal volume of culture medium which is MEM culture medium of 5% fetal calf serum, and repeating for 3 times.

(4) Placing at 37 deg.C with 5% CO₂The culture box is continuously cultured, the culture medium is removed after 48 hours of culture, PBS is used for washing for 3 times, the total RNA of samples of each hole is extracted, and the RT-PCR is used for detecting the propagation of each test medicament to Eimeria tenella (E.tenella)The influence of (c).

The Real-time PCR primers were as follows:

specifically amplifying Eimeria tenella (E.tenella) actin primer,

EtActin-F:5’-CACCACCGCCGAGAAAGA-3’(SEQ ID NO.1)，

EtActin-R:5’-GAACAACATTG-CCGTAGAGG-3’(SEQ ID NO.2)；

specific amplification host cell actin primer,

Bactin-F:5’-GGATGAGGCTCAGAGCAAGAGA-3’(SEQ ID NO.3)，

Bactin-R:5’-TCGTCCCAGTTGGTG-ACGAT-3’(SEQ ID NO.4)。

by Delta C_TMethods the effect of each test drug on the growth and development of eimeria tenella (e.tenella) at the cell culture level was calculated.

The Real-time PCR reaction system is shown in Table 1, and the Real-time PCR reaction program is shown in Table 2.

TABLE 1 Real-time RT-PCR reaction System

TABLE 2 Real-time PCR reaction procedure

Evaluation indexes are as follows: delta C of experimental and control groups was obtained by Real-time RT-PCR_T＝C_T[Et actin]-C_T[H actin]，ΔΔC_T＝ΔC_T[exp]-ΔC_T[ref]Finally, the effect of resisting the Eimeria tenella (E.tenella) of the medicament to be screened is obtained:

thirdly, experimental results:

respectively calculating new Zealand vitexin 2 groups,The inhibitory effect of the drugs of the New Zealand vitexin 3 group and the positive control group on the Eimeria tenella (E.tenella), the inhibitory effects of the New Zealand vitexin 2, the New Zealand vitexin 3 and the control drug sulfachloropyrazine sodium on the reproduction of the Eimeria tenella (E.tenella) are obvious (Table 3), and the New Zealand vitexin 2 and the New Zealand vitexin 3 on the MIC of the Eimeria tenella (E.tenella)₅₀Values were 28.35. mu.M, 35.08. mu.M, respectively, MIC of the control drug sulfachloropyrazine sodium for Eimeria tenella (E.tenella)₅₀The value was 46.61. mu.M. The result shows that both the vitexin 2 and the vitexin 3 have good anti-eimeria tenella effect and are obviously superior to the sulfachloropyrazine sodium serving as a reference medicament.

TABLE 3 inhibition of coccidial reproduction at cellular level by each test drug

FIG. 1 is a graph showing the effect of Vitexin 2 in New Zealand on the reproduction of Eimeria tenella; in the figure, the abscissa is the concentration of compound vitexin 2, and the ordinate is the inhibition rate of compound vitexin 2 against the propagation of eimeria tenella; FIG. 2 is a graph showing the effect of Vitexin 3 in New Zealand on the reproduction of Eimeria tenella; in the figure, the abscissa is the concentration of compound vitexin 3, and the ordinate is the inhibition rate of compound vitexin 3 against the propagation of eimeria tenella; FIG. 3 is a graph of the effect of sulfachloropyrazine sodium on the propagation of Eimeria tenella; in the figure, the abscissa is the concentration of the compound sulfachloropyrazine sodium, and the ordinate is the inhibition rate of the compound sulfachloropyrazine sodium on the propagation of Eimeria tenella. As can be seen from fig. 1 to 2, as the concentrations of zealand vitexin 2 and 3 are increased, the effect of zealand vitexin 2 and 3 on inhibiting the proliferation of eimeria tenella is enhanced, the effect of zealand vitexin 2 and 3 on inhibiting the proliferation of eimeria tenella has concentration dependency, and the activity of zealand vitexin 2 and 3 on inhibiting the development and proliferation of eimeria tenella (e.tenella) is good.

Example 2

This example provides animal level assays for New Zealand Vitexin 2 and New Zealand Vitexin 3 as follows:

firstly, experimental materials:

(1) preparation of a new zealand vitexin 2 powder preparation formulation: respectively mixing 1g of New Zealand vitexin 2 (not less than 80%) and 99g of starch uniformly; preparation of a new zealand vitexin 3 powder preparation: mixing 1g of New Zealand vitexin 3 (not less than 80%) and 99g of starch respectively.

(2) Control drugs: a conventional anticoccidial drug, Triplex coccidium powder (30% sulfachloropyrazine sodium), was produced by Norwalk animal health Limited, Shanghai.

(3) Coccidian oocysts: eimeria tenella (e.tenella) eurycosporulated oocysts were stored by the institute of animal health, academy of agricultural sciences, guangdong, and rejuvenated in coccidiless chicks before use.

(4) Chicks: lingnan yellow broiler chickens are provided by animal science research institute of Guangdong province academy of agricultural sciences and are raised in a sterilized special animal house; the chicken coop and the utensils are strictly disinfected, and the chicken coop can freely eat and drink purified water; before the test, the chicks are observed to have clinical symptoms and the feces are continuously checked for coccidian oocysts for later use for 2 days.

(5) Feed: the young feed is prepared by Xinnandu feed science and technology Limited company in Guangdong province, and does not contain any anticoccidial drugs.

II, an experimental method:

(1) grouping: 180 south-south yellow broilers are raised to 10 days old according to test grouping and processing, weighing one by one, eliminating weak chicks and overweight chicks, randomly dividing the rest chicks into 5 groups of 30 chicks, and properly adjusting to ensure that the total weight of each group of chicks is approximately equal.

(2) And (3) treatment: except for the normal daily feed of the group 1 (blank control), the chicks of the groups 2 to 5 were orally administered with E.tenella (E.tenella) sporulated oocysts lx 10⁵Individuals/feather; on the day of attack, 3 rd to 5 th groups of the chickens are respectively fed with feed containing the coccidiostat for 7 days, and the grouping condition of the test chickens and the usage and dosage of the feed are shown in table 4.

TABLE 4 Experimental grouping

(3) And (4) counting results:

observing and recording the mental state, feed intake, excrement condition and the like of the chicken flocks every day; weighing dead chicks, performing a autopsy, and scoring the lesion as +4 points if the chicks die due to Eimeria tenella (E.tenella) infection; weighing all chicks one by one on the 7 th day after infection, performing a dissection, and scoring for caecum lesion; and finally, calculating the weight gain and feed reward of each group of chicks.

The efficacy determination method and the standard anticoccidial index (ACI) were calculated according to the formula recommended by merck:

ACI ═ (relative rate of weight gain + survival) - (oocyst value + lesion value);

relative percent (%) gain (test group weight gain ÷ blank control group weight gain) × 100%;

survival (%) — (number of surviving chicks per group ÷ total number of chicks per group) × 100%.

Lesions were scored on a five-point scale:

firstly, no oocysts exist, and the caecum is normal and is divided into 0 point;

secondly, oocysts exist, the mucous membrane of the caecum is slightly thickened, and a small amount of intestinal contents scattered in bleeding or a small amount of blood sample is divided into +1 point;

③ 2 minutes for oocyst, thickening of caecum mucous membrane and obvious bleeding or obvious blood sample intestinal content;

fourthly, oocysts exist, mucous membranes of caecum thicken, and a large amount of blood clots or blood sample intestinal cores are present, and the score is + 3;

chicks died due to coccidiosis or had a large number of oocysts, the cecum appeared dark reddish brown (or the middle of the small intestine had punctate necrotic foci, the mucous membrane appeared crimson), the intestinal canal was enlarged significantly, and the content formed a significant blood sample intestinal core, which was +4 points.

Mean lesion score per group x 10.

Oocyst values are converted from number of oocysts per gram of fecal contents (OPG).

The drug effect judgment standard is as follows: ACI <120 is ineffective, 120-160 is weak effect, 160-180 is middle effect, more than 180 is strong effect.

Thirdly, experimental results:

(1) observation of clinical symptoms: on day 2 of the control group (group 2) inoculated with sporulated oocysts of eimeria tenella (e.tenella), test chickens developed depressed spirit and decreased food intake, and developed a clear bloody stool and died on day 5, with the bloody stool gradually decreasing thereafter. Groups 3, 4 and 5 had a slight decrease in appetite on day 3 after inoculation with Eimeria tenella sporulated oocysts. The diet and mental status of the test chickens in group 1 were normal during the experiment.

(2) Anticoccidial indexes of groups 1 to 5 were calculated, respectively, and the results are shown in Table 5. The anticoccidial indexes of the groups 4 and 5 (the group 2 and the group 3) are higher than those of the group 3 (three-character coccidium powder) of the control medicament, and the anticoccidial indexes (ACI) of the groups 4 and 5 can reach more than 165, so that the anticoccidial index has good effect of resisting the eimeria tenella (E.tenella), and is superior to the three-character coccidium powder of the conventional anticoccidial medicament.

TABLE 5 anticoccidial index of each group

In summary, the embodiment of the invention discloses that the New Zealand vitexin 2 and the New Zealand vitexin 3 can be used for treating infection of Eimeria tenella, the application of the New Zealand vitexin 2 and the New Zealand vitexin 3 is expanded, and a new anti-coccidium drug is developed. The invention carries out cell level tests and animal tests, and the results show that the New Zealand vitexin 2 and the New Zealand vitexin 3 have obvious effect of inhibiting Eimeria tenella. The new zealand vitexin 2 and the new zealand vitexin 3 provided by the invention can be used for preparing an anti-eimeria tenella medicine.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

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Claims

1. Use of at least one of New Zealand vitexin 2 and New Zealand vitexin 3 in preparing medicine or feed for resisting coccidiosis.

2. Use of at least one of vitexin 2 and vitexin 3 of New Zealand according to claim 1 for the preparation of a medicament or feed against coccidiosis, wherein the coccidiosis is chicken coccidiosis.

3. Use of at least one of vitexin 2 and vitexin 3 of new zealand in the preparation of a medicament or feed against coccidiosis according to claim 2, wherein the infectious protozoa of coccidiosis in chicken is eimeria tenella.

4. The use of at least one of vitexin 2 and vitexin 3 of New Zealand according to claim 2 in the preparation of a medicament or feed for combating coccidiosis, wherein the chicken variety infected with said coccidiosis is Lingnan yellow broilers.

5. Use of at least one of vitexin 2 and vitexin 3 of New Zealand in the preparation of a coccidiosis-resistant medicament or feed according to any one of claims 1 to 4, wherein the medicament comprises at least one of vitexin 2 and vitexin 3 of New Zealand and pharmaceutically acceptable auxiliary materials.

6. Use of at least one of vitexin 2 and vitexin 3 according to any one of claims 1 to 4 in the preparation of a medicament or feed for resisting coccidiosis, wherein the medicament is administered by a herd.

7. Use of at least one of vitexin 2 and 3 according to any one of claims 1 to 4 in preparation of a coccidiosis-resistant medicament or feed, wherein the medicament is in the form of powder.

8. Use of at least one of vitexin 2 and 3 according to any one of claims 1 to 4 in preparation of anti-coccidiosis drugs or feeds, wherein the drug is in the form of water-soluble powder.

9. The use of at least one of vitexin 2 and 3 in the preparation of anti-coccidiosis drugs or feeds according to any one of claims 1 to 4, wherein the drug is in the form of a solution.

10. The use of at least one of vitexin 2 and 3 according to any one of claims 1 to 4 in the preparation of a coccidiosis-resistant medicament or feed, wherein the medicament is in the form of microcapsules.

11. Use of at least one of vitexin 2 and 3 according to any one of claims 1 to 4 in the preparation of a coccidiosis-resistant medicament or feed, wherein the medicament is in the form of a tablet.

12. Use of at least one of vitexin 2 and 3 according to any one of claims 1 to 4 in the preparation of a medicament or feed for combating coccidiosis, wherein the feed is a complete formula feed, a concentrated feed, a premix/premix feed, a concentrate mixture or a mixed feed.